Enhanced antimicrobial activity compositions of blends of plant essential oils

ABSTRACT

Antimicrobial compositions based on a combination or blend of plant essential oils is of enhanced antimicrobial effectiveness; by adding to the combination of at least two plant essential oils, and preferably adds a small but antimicrobial enhancing effective amount of an enhancer selected from the group consisting of polyionic organic enhancers and polyionic inorganic enhancers. One preferred blended oil composition is a mixture of plant essential oils wherein at least one of the oils is oregano oil. The oil blend is used as a major component in the finished product anti-microbial.

CROSS-REFERENCE TO RELATED APPLICATIONS

This Continuation-in-part application claims priority under 35 U.S.C.§120 of application Ser. No. 12/426,622 filed Apr. 20, 2009 nowabandoned, and under 35 U.S.C. §119(e) to provisional application Ser.No. 61/053,216 filed May 14, 2008, each of which are herein incorporatedby reference in their entirety.

FIELD OF THE INVENTION

This invention relates to methods and compositions using plant essentialoil blends as antimicrobials.

BACKGROUND OF THE INVENTION

It is known in the art that plant essential oils, that is oils derivedfrom plants by distillation, expression or extraction may haveantimicrobial activity when exposed to bacterial cells, yeast and mold.Consumer acceptance of these essential oils is high because they usuallyhave the pleasant fragrance of the plant from which they were extracted.When used for animal use, the animals do not commonly shy away from thembecause again, they have the odor of plants from which they werederived, and such odors are not unfamiliar to many animals.

Essential oils mixed with carriers have a lot of potential veterinaryand human uses. For example in the veterinary world they may be used asteat dips, or disinfecting topicals such as for skin ulcers, forshampoos, for topical gels and creams, for anti-fungals, and even can betaken internally for use in the GI tract, such as for scours products.These final or finished products are collectively referred to herein as“finished products”.

There is a continuing need for increasing the cellular uptake of plantessential oils in order to enhance their anti-bacterial effect. Someresearchers have theorized that plant essential oils soften the walls ofthe bacteria then permeate them thus causing an enhanced anti-bacterialeffect. (see, Vaara, “Agents That Increase the Permeability of the OuterMembrane”, Microbiological Reviews, September 1992, Vol. 56(3); andJohnson U.S. Pat. No. 6,319,958 that teaches addition of at least onesesquiterpenoid to advance the antimicrobial effect of antimicrobialcompounds.

In the ever increasing efforts to enhance antimicrobial effectivenessApplicant has now discovered that a combination of plant essential oils(blends) provides increased enhancement; and moreover the combination ofoils may be used with other known additional enhancers to even furthermaximize effectiveness of many disinfecting topicals.

Accordingly it is an object of the present invention to provide plantessential oil-derived antimicrobial compositions, that use a combinationof essential oils, and in some cases added enhancers are also used withthe oil blends, to achieve a maximized antimicrobial effect.

It is also an object of this invention to prepare a variety of differentantimicrobial compositions based on the above discovery that are usefulfor veterinary or human use.

The method or means of accomplishing at least the above objectives willbecome apparent from the detailed description of the invention whichfollows hereinafter.

BRIEF SUMMARY OF THE INVENTION

Antimicrobial compositions based on a combination of plant essentialoils are of enhanced antimicrobial effectiveness and are prepared byadding a blend of at least two plant essential oils preferably with asmall but antimicrobial enhancing effective amount of an additionalenhancer selected from the group consisting of polyionic organicenhancers and polyionic inorganic enhancers to a disinfectingcomposition such as a teat dip or scours treatment. One preferredcombination or blend is a mixture of plant essential oils wherein atleast one of the oils is oregano oil.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

This invention relates to antimicrobial compositions derived from plantessential oils, more importantly a combination of essential oils toenhance antimicrobial effectiveness. The essential oil component as aconcentrate may be from 40% to 95% by weight oils, but is preferablyfrom 50% to 90% by weight of a combination of essential oils. Thebalance is usually a known organic acid anti-microbial such as citricacid, butyric acid, fumaric acid, lactic acid and acetic acid. The mostpreferred essential oils being a 1 to 1 by weight mix of at least twodifferent essential oils, with one of them being oregano oil. As a majorcomponent as herein defined means of the total concentrate at least 40%by weight.

Essential oils are volatile aromatic oils which may be synthetic or maybe derived from plants by distillation, expression or extraction, andwhich usually carry the odor or flavor of the plant from which they areobtained. In the combination compositions of this invention, antisepticactivity is provided by essential oils. Some of these essential oilsalso act as flavoring agents. Besides oregano oil and thymol, theessential oils of this invention may include but are not limited tomenthol, methyl salicylate (wintergreen oil), eucalyptol, carvacrol,camphor, chamomile, anethole, carvone, eugenol, isoeugenol, limonene,osimen, n-decyl alcohol, citronel, a-salpineol, methyl acetate,citronellyl acetate, methyl eugenol, cineol, linalool, ethyl linalaol,safrola vanillin, spearmint oil, peppermint oil, lemon oil, orange oil,sage oil, rosemary oil, cinnamon oil, pimento oil, laurel oil, cedarleaf oil, and clove oil.

In embodiments of the invention wherein organic phenolic compounds areobtained from plant oil extracts, the oil is preferably extracted from amember of the Labiatae (also called Lamiaceae) or Verbenaceae family.Plants in the family Labiatae or Verbenaceae include hybrids of plantsproduced from individual plants in those two families.

The common name for members of the Labiatae family, a large family ofmostly annual or perennial herbs, is the “mint family.” The mint familyis classified in the division Magnoliphyta, class Magnoliopsida, andorder Lamiales. The Labiatae family includes about 200 genera, such asSalvia, Rosmarinus, Mentha, Ocimum, Thymus, Marrubium, Monarda,Trichostema, Teucrium, Hyptis, Physostegia, Lamium, Stachys, Scutellariaand Lycopus.

Plants which are preferably used for extraction of organic phenoliccompounds include, but are not limited to, Ocimum spp., Saturea spp.,Monarda spp, Origanum spp, Thymus spp., Mentha spp., Nepeta spp.,Teucrium gnaphalodes, Teucrium polium, Teucrim divaricatum, Teucrimkotschyanum, Micromeria myrifolia, Calamintha nepeta, Rosmarinusofficinalis, Myrtus communis, Acinos suaveolens, Dictamnus albus,Micromeria fruticosa, Cunila origanoides, Mosla Japonoica Maxymowitz,Pycnanthemum nudum, Micromeria Juliana, Piper betel, Trachyspermum ammi,Lippia graveolens Escholcia splendens, and Cedrelopsis grevei, as wellas others.

In a preferred composition, the oil is extracted from Esholtiasplendens, Cedrelopsis grevei, Lippia graveolens or a plant of thespecies Nepeta, including but not limited to Nepeta racemosa (catmint),Nepeta citriodora, Nepeta elliptica, Nepeta hindostoma, Nepetalanceolata, Nepeta leucophylla, Nepeta longiobracteata, Nepeta mussinii,Nepeta nepetella, Nepeta sibthorpii, Nepeta subsessilis and Nepetatuberosa.

Most preferably, the oil is extracted from a hybrid plant produced fromcrossing Nepeta racemosa, Esholtia splendens, Cedrelopsis grevei, andLippia graveolens.

Plants of the Labiatae and Verbenacea families are found throughout theworld and are relatively easy to cultivate. To cultivate the plants,seeds, preferably those of plants that are expected to yield a highpercentage (e.g., at least about 70 wt %, more preferably at least about80 wt %), of organic phenolic compounds, are planted in fine loose soil,preferably in a sub-tropical climate. Hybrid seeds having a highpercentage of organic phenolic compounds can be produced by knowntechniques. Crossing Nepeta racemosa, Esholtia splendens, Cedrelopsisgrevei, and Lippia graveolens produces one such hybrid that is apreferred source of the organic phenolic compounds. The seeds are thencultivated using known agricultural techniques, such as watering, andartificial fertilizing. Most preferably, the plants are cultivated andgrown without the use of any synthetic pesticides.

Because the leaves contain a high amount of oil upon blossoming, it ispreferred that the plants be harvested soon after the plants begin toblossom. Preferably, the plants are harvested within 24 hours afterblossoming, more preferably within 12 hours after blossoming. Mostpreferably, harvesting is undertaken early in the morning or late in theevening hours (after blossoming begins) when the leaves are not exposedto the sun.

Because the majority of the oil is found in the leaves and blossoms ofthe plant, it is preferred that only the leaves and blossoms be utilizedin the extraction process. Use of other parts of the plant may increaseimpurities and decrease yield, but may be utilized.

Thymol, also known by the chemical formula 5-methyl-2-(1-methylethyl)phenol, is obtained from the essential oil of Thymus vulgaris Labiataeand Monarda punctata Labiatae. Thymol is a white crystalline powder withan aromatic odor and taste and is soluble in organic solvents but onlyslightly soluble in deionized water. Thymol along with oregano oil arepreferred for at least one of the combination of essential oils

Menthol is isolated principally from the oil of Mentha arvensis. In itscommercial form, menthol is available as L-menthol crystals obtainedfrom a process involving cooling of the oil. Fractional distillation ofpeppermint oil which usually contains from about 40% to about 65%menthol represent another important source of menthol. Synthetic sourcesof L-menthol are also available.

Eucalyptol, another essential oil with antiseptic properties, is derivedfrom the eucalyptus tree. Having a camphoraceous odor and cooling taste,this essential oil is often combined with other essential oils such asmenthol in confection formulations to impart medicinal effect.Combinations of menthol and eucalyptol are widely used. Particularlypreferred uses of the menthol-eucalyptol combination include, accordingto the present invention, dentifrices such as toothpastes or dentalgels.

Methyl salicylate is the main ingredient in many essential oils,constituting about 99% of oil of wintergreen (Gaultheria procumbens) andsweet birch (Betula lento). Methyl salicylate, which has a distinctiverefreshing aroma, is used widely in mouthwashes, chewing gums and otheroral and pharmaceutical preparations.

In the most preferred compositions of the present invention it ispreferred that at least one of the essential oils have as their activeingredient a combination of thymol and carvacrol. The most preferred isoregano oil.

One very satisfactory oil blend is 47.5% by weight oregano oil, 23.75%by weight cinnamon bark oil, and 23.75% by weight clove oil and 5%capsicum oil resin. Another oil blend which may also be used is: 46% byweight oregano, 22% cinnamon bark, 22% clove, 5% nerolidol and 5%capsicum.

A third blend formulation is 30% oregano, 30% cinnamon bark, 30% clove,5% nerolidol and 5% capsicum.

A fourth blend formulation is 36.20% oregano, 18% cinnamon bark, 17%clove, 4% nerolidol, 0.8% oleoresin capsicum, 4% cranberry, 6.60%geranium, 6.67% patchouli, and 6.67% tea tree.

A fifth blend formulation is 33% regular oregano, 33.34% clove, and33.34 cinnamon.

A sixth blend formulation is 95% rosemary oil and 5% nerolidol.

The essential oils can be mixed in a variety of physical formats, withone preferred one being so called beads. Bead format is from 0.5% to 50%of a combination of oils or pure oil added to a mixture of alginate,shellac and seaweed carriers to provide a carrier bead with the oil oroils. This allows for a convenient and easy subsequent processing. Thebeading process is known and can be accomplished by a variety ofmanufacturers.

Products based on essential oils, such as those containing organicphenolic compounds, tend to be absorbed at a level greater than 90% inthe small intestines. Therefore, most of the activity of such productstends to be localized in the stomach and/or small intestine. However,there are many microbial infections that occupy portions of thegastrointestinal tract beyond the small intestine. Therefore, it may bedesirable to extend the activity of the combination essential oil basedproduct into the large intestine.

Microencapsulation is one method that can help extend the-activity ofthe antimicrobial composition throughout the entire gastro-intestinaltract (GIT). Microencapsulation is a micro-packaging technique whichinvolves the coating of small particles of solids, liquid droplets, ordispersion of solids, within liquids. Microencapsulated antimicrobialcompound may be used to treat infections located in the end of the smallintestines (e.g., jejunum and/or ileum) and beginning of the largeintestines (e.g., ascending colon and transverse colon). Themicroencapsulation prevents release of the active ingredients in thestomach or in the beginning of the small intestines (e.g., duodenum). Ifthe antimicrobial compound is not microencapsulated, the acidicenvironment of the stomach will tend to break the association betweenthe antimicrobial compound and most carriers in the pharmaceuticalcomposition (such as dextrose, starch, etc.) and thereby activate theantimicrobial compound in the stomach.

For example, a microencapsulated form of the antimicrobial compositionsmay be used to treat Cryptosporidia spp. infections and/or chronicenteritis in humans; Cryptosporidia infections in animals, Lawsoniaintracellularis and Treponema hyodesynteriae infections in pigs, andothers.

One example of a microencapsulation process includes encapsulating theantimicrobial composition in a multi walled capsule such that the layersof the wall dissolve as the capsule travels through the gastrointestinaltract. Thus, the components that make up each layer of the capsule wallare chosen based on the conditions in the specific region of thegastrointestinal tract in which they are desired to dissolve. Forexample, the pH along the gastrointestinal tract (GIT) varies: in thestomach, the pH is between 2 and 5; in the duodenum, 4 and 6; jejunum, 4and 6; ileum, 6.5 and 7.5; caecum 5.5 and 6.5; colon, 6.5 and 7; andrectum, 6.5 and 7. Therefore, the components of the wall layers maydiffer depending on what type of an ailment is to be treated, or itslocation, and whether the final formulation is meant to treat humans oranimals. Each layer of the wall may also contain the composition of theinvention so that upon dissolution of that wall layer, it can bereleased to effectuate treatment of the ailment.

Suitable coating matrices include fatty acids, waxes, sugars, andshellac.

Encapsulation techniques are known. An example of one encapsulationtechnique (called fluidized bed coating) is provided below. In afluidized bed, a suspension of solid particles is transformed into afluid-like state by an upward gas flow through the system. Because ofthe intensive heat and mass transfer, fluidized bed reactors are widelyused, e.g. in chemical industry for solid-catalyzed gas-phase reactions.To maximize the yield of such reactors, liquid reactants can locally beinjected into the fluidized bed. The injected liquid reactants penetratethe fluidized bed and evaporate. For design purposes and the achievementof optimal operating conditions, the spatial distribution of theconcentration of components and temperature has to be predicted.

Fluidized bed coating can be used to encapsulate the antimicrobialcompound in a coating material which includes ethyl cellulose and plantoil. First, the antimicrobial compound described is combined in thefluid bed mixer with the ingredients to form a powder, such as theingredients shown in the table below.

While the above disclosure has been emphasizing blends of at least twooils with at least one being oregano, multi-oil blends with at least onebeing oregano also work.

CONCENTRATE EXAMPLES

Three especially preferred oil blend formulas include the following:

1^(st) Oil Blend Formula: Base Formula* 33.34% Regular Oregano Oil33.34% Rosemary Oil 11.11% Licorice Powder 11.11% Cinnamon Bark 11.11%  100% 2^(nd) Oil Blend Formula: Base Formula* 33.34% Regular OreganoOil 33.34% Rosemary Oil 11.11% Chamomile Oil 11.11% Peppermint Oil (highmenthol 11.11% content)   100% 3^(rd) Oil Blend Formula: Base Formula*33.34% Regular Oregano Oil 33.34% Cinnamon Bark Oil 11.11% Rosemary Oil11.11% Peppermint Oil (high menthol 11.11% content)   100% These may beused with additional enhancers and other ingredients to form finishedproducts. *Base Formula as used here includes 33% Geranium oil, 33.34%Patchouli and 33.34 Tea Tree oil.

SCOURS FINISHED PRODUCT EXAMPLE

Two scours treating products were made, each containing combinations ofessential oils, as listed below in the finished product. Product Acontained sodium polyphosphate inorganic enhancer. Product B containedthree enhancers Nerolidol, PEI, and sodium polyphosphate.

A B Ingredient Percentage Ingredient Percentage Purified Water 44.60%Purified Water 39.60% Activated Charcoal 10.00% Activated Charcoal10.00% Regular Oregano Oil 7.50% Regular Oregano Oil 7.20% Cinnamon BarkOil 3.25% Cinnamon Bark Oil 7.20% Redistilled Clove 2.75% RedistilledClove 3.60% Leaf Oil Leaf Oil Nerolidol 0.50% Nerolidol 1.00% CapsicumOleoresin 0.75% PEI 0.50% Cranberry Extract 0.25% Sodium Polyphosphate0.50% Dextrose 5.00% Dextrose 5.00% Agro-Pect 5.00% Agro-Pect 5.00%Arabic Gum 2.50% Arabic Gum 2.50% Glycine 2.50% Glycine 2.50% PsyliumSeed Hulls 2.50% Psylium Seed Hulls 2.50% Betaine Hydrochloride 2.00%Betaine Hydrochloride 2.00% Seaweed Meal---Nori 2.00% SeaweedMeal---Nori 2.00% Flakes Flakes Citric Acid 1.50% Citric Acid 1.50%Sodium Chloride (Extra 1.50% Sodium Chloride 1.50% Fine) Salt (ExtraFine) Salt Potassium Chloride 1.50% Potassium Chloride 1.50% (Muriate ofPotash) (Muriate of Potash) Magnesium Chloride 1.00% Magnesium Chloride1.00% Citricidal 1.00% Citricidal 1.00% Ascorbic Acid 1.00% AscorbicAcid 1.00% Zinc Oxide 1.00% Zinc Oxide 1.00% Magnesium Oxide 0.30%Magnesium Oxide 0.30% Chlorophyll 0.10% Chlorophyll 0.10%

Each of products A and B when mixed appeared physically as a geldelivery format. In several separate trials over 1000 baby calves weretreated in the following manner. The gel is orally fed to the strickencalf at 30 cc/calf as a 1-3 time dose depending upon the severity ofdiarrhea. In nearly all cases after treatment the scours cleared upwithin days.

As used in the appended claim with respect to the oil blend concentrate,major component refers to at least 40% by weight of the totalconcentrate being the blended essential oil component; minors includeother additives.

A polymeric polyionic organic enhancer can be added to the concentrateand can be the preferred polyethyleneimine (PEI) or can be others suchas paramethoxyphenyl ethylmethylamine. The amount can be 0.1 mM to 50mM, similar to the same amount of the sesquiterpenoids (herein beforedescribed).

The polyionic inorganic enhancers if used are preferably polyphosphateenhancers and can include sodium tripolyphosphate, sodiumhexametaphosphate, at similar levels.

Other carriers may include minors used for a variety of purposes invarious topicals, pills, gelatins, etc. and can include small amounts ofApple Powder, Citrus Pectins, Arabic Gum, Ascorbic Acid, Beeswax,Betaine Hydrochloride, Biotin, Calcium Carbonate (Thermocal), CanolaOil, Cetyl Alcohol, Choline Chloride, Citric Acid, Cobalt Carbonate,Copper Sulfate, Corn Starch, Dextrose, Dry Sweet Orange Flavoring,Flaxseed Oil, Folic Acid, Glycine, Lanolin, Lavendar Oil, Lemon Powder,Lipase DS, Maltodextrin, Manganese Sulfate, Magnesium Chloride,Magnesium Oxide, Malic Acid, Niacin, Olive Oil, Pantothenic Acid,Potassium Chloride, Potassium Sulfate, Polysorbate, Propylene Glycol,Purple Pigment, Pyridoxine HCL, Riboflavin, Seaweed Meal,Probiotics/Bacteria, Selenium, Silicon 505, Silicon Dioxide, SodiumAcetate, Sodium Chloride, Sodium Citrate, Sodium Propinate, SodiumSilica Aluminate—MS, Spearmint Oil, SST (Activated Charcaol), SterylAlcohol, Thiamine, Vaseline, Vitamin A, Vitamin B12 600 mg, Vitamin D3500, Vitamin E, Vitamin K, Water, and Zinc Sulfate.

Applicants have also discovered that further anti-microbial propertyenhancement is achieved if from 0.01% by weight to 10% by weight,preferably 0.05% by weight to 5.0% by weight of the concentrate is anorganic acid anti-microbial selected from the group of acetic, butyric,citric, lactic and fumaric.

Testing of the combination of oils at Iowa State University during theyear 2006/2007 revealed that the combination of oils was more effectivefrom an antimicrobial standpoint than the single oils alone. Preferredwas the oil blend previously described as the first oil blend and thesecond oil blend. These oils in combination with various carriers inconcentrate form may be used to add to a variety of veterinarian andhuman use finished product compositions; and, those can include indelivery format pills, gelatin capsules, skin topicals, gels, creams,liquid rub-ons, powders, shampoos, G.I. tract medicines, etc.

To make a final finished product the concentrate containing the plantessential oil blend is added to the remaining ingredients of thefinished product to provide from 0.05% to about 25% by weight of theoverall finished product, preferably from 0.5% to 10% by weight of thefinished product, and most preferably from 0.5% to 5% by weight of thefinished product.

The combination of essential oils, and the enhancers offer manyadvantages for the anti-microbial composition of finished products. Forone, they are not widely rejected by treatment animals because theycontain natural scents of the plants from which they are derived, andthe animals are used to these scents. Another advantage is the enhancedeffectiveness as an anti-microbial. A third is the ease ofprocessability with other finished product ingredients. And a stillfurther advantage is the wide variety of delivery formats that can bepresented.

It therefore can be seen that the invention accomplishes at least all ofits stated objectives.

1. A method of increasing the cellular uptake of plant essential oils incells comprising: exposing the cells to a combination of: a) anantimicrobial effective amount of at least two essential oils includingoregano oil and cinnamon oil; and b) an effective amount of cellularuptake enhancer selected from the group consisting of polyorganic ionicenhancer, and polyinorganic ionic enhancers.
 2. The method of claim 1wherein the combination of plant essential oils further includes thymol.3. The method of claim 1 wherein the combination of essential oils andenhancer are microencapsulated in a multi-walled capsule.
 4. The methodof claim 3 wherein the components of the multi-walled capsule are chosenbased upon the specific region of the gastrointestinal tract in whichthey are desired to dissolve.